Tiwari Ananda, Paakkanen Jaana, Österblad Monica, Kirveskari Juha, Hendriksen Rene S, Heikinheimo Annamari
Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland.
Expert Microbiology Unit, Finnish Institute for Health and Welfare, Kuopio, Finland.
Front Microbiol. 2022 Jun 2;13:887888. doi: 10.3389/fmicb.2022.887888. eCollection 2022.
Antimicrobial resistance profiling of pathogens helps to identify the emergence of rare or new resistance threats and prioritize possible actions to be taken against them. The analysis of wastewater (WW) can reveal the circulation of antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARG) among the catchment communities. Here, we analyzed WW influent samples to determine the prevalence of carbapenemase genes-carrying Gram-negative bacteria (Carba-GNB) in Helsinki, Finland. This study set important historical reference points from the very early stage of the carbapenemase era, during the period 2011-2012. A total of 405 bacterial isolates grown on CHROMagarKPC ( = 195) and CHROMagarESBL ( = 210) from WW influent samples were collected between October 2011 and August 2012 and were analyzed. The bacterial DNA from the isolates was extracted, and the prevalence of carbapenemases genes , , , , , , and were screened with multiplexed PCR. All carbapenemase-positive isolates were identified taxonomically to species or genus level with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The nucleic acid extraction was successful for 399 isolates, of which 59 (14.8%) were found to carry carbapenemase genes. A total of 89.8% of the carbapenemase positive isolates (53 out of 59) were obtained from CHROMagarKPC plates and only 10.2% (six out of 59) were obtained from CHROMagar ESBL plates. Among the Carba-GNB isolates, 86.4% were (51 out of 59), 10.2% were (six out of 59), and 3.4% were (two out of 59). The most common carba-gene, , was carried by 10 different bacterial species, including spp., spp., and spp.; the gene was carried by , , and ; and the gene was carried by / and . This study emphasizes that wastewater surveillance (WWS) can be an additional tool for monitoring antimicrobial resistance (AMR) at the population level.
对病原体进行抗菌药物耐药性分析有助于识别罕见或新出现的耐药威胁,并确定针对这些威胁可能采取的行动的优先级。废水(WW)分析可以揭示集水区社区中抗菌药物耐药菌(ARB)和抗菌药物耐药基因(ARG)的传播情况。在此,我们分析了芬兰赫尔辛基废水进水样本,以确定携带碳青霉烯酶基因的革兰氏阴性菌(Carba-GNB)的流行情况。本研究在2011 - 2012年期间,从碳青霉烯酶时代的早期阶段设定了重要的历史参考点。2011年10月至2012年8月期间,共收集了405株在CHROMagarKPC(= 195)和CHROMagarESBL(= 210)上生长的来自废水进水样本的细菌分离株并进行分析。提取分离株的细菌DNA,并用多重PCR筛选碳青霉烯酶基因blaKPC、blaNDM、blaVIM、blaOXA-48、blaOXA-23、blaOXA-58、blaIMP和blaGES的流行情况。所有碳青霉烯酶阳性分离株通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)在种或属水平上进行分类鉴定。399株分离株的核酸提取成功,其中59株(14.8%)被发现携带碳青霉烯酶基因。总共89.8%的碳青霉烯酶阳性分离株(59株中的53株)来自CHROMagarKPC平板,只有10.2%(59株中的6株)来自CHROMagar ESBL平板。在Carba-GNB分离株中,86.4%是肺炎克雷伯菌(59株中的51株),10.2%是大肠埃希菌(59株中的6株),3.4%是鲍曼不动杆菌(59株中的2株)。最常见的碳青霉烯酶基因blaKPC由10种不同细菌携带,包括肺炎克雷伯菌、大肠埃希菌和鲍曼不动杆菌;blaNDM基因由阴沟肠杆菌、弗劳地枸橼酸杆菌和产气肠杆菌携带;blaVIM基因由嗜麦芽窄食单胞菌/栖稻假单胞菌携带。本研究强调废水监测(WWS)可以成为在人群水平上监测抗菌药物耐药性(AMR)的另一种工具。